CN110721174B - 一种用于防治鱼类粘孢子虫病的药物 - Google Patents
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Abstract
本发明涉及一种用于防治鱼类粘孢子虫病的药物，属于水产养殖技术领域。其提供一种用于防治鱼类粘孢子虫病的药物，包括香芹酚。所述香芹酚的浓度范围以鱼体重计算，为50‑500 mg/kg。本发明提供的药物在离体条件下能有效杀死粘孢子虫成熟孢子，添加到饲料中投喂能有效预防控制鱼类粘孢子虫病发生。
Description
技术领域
本发明涉及一种用于防治鱼类粘孢子虫病的药物，属于水产养殖技术领域。
背景技术
粘孢子虫是一类适应宿主体内寄生生活、个体微小的刺胞动物，主要感染海淡水鱼类及两栖动物，已报道约2500多种；其中一些种类会造成养殖或自然水体鱼类严重疾病和大量死亡，如洪湖碘泡虫（Myxobolus honghuensis）引起的异育银鲫咽部肿大、溃烂，脑碘泡虫(Myxobolus cerebralis)引起的鲑鳟鱼“旋转病”，鲢碘泡虫(Myxobolus drjagini)引起的“鲢疯狂病”，吉陶单极虫（Thelohanellus kitauei）引起的鲤肠道“巨囊病”等。由于寄生虫生活史复杂性和发育阶段的多样性，粘孢子虫研究一直缺乏实验室条件下稳定的传代保存和感染模型，严重限制了病原致病机制、治疗药物缺乏科学有效的筛选和评价方法。生产中防治鱼类粘孢子虫病的常用药物，如青蒿素、妥曲珠利、盐酸氯苯胍和敌百虫等，多数借鉴防治人类疟原虫和畜禽球孢虫的药物。粘孢子虫与疟原虫、球孢虫在分类进化上差异非常大，目前常用的药物杀灭粘孢子虫的效果不明确，甚至无效。
香芹酚(carvacrol，2-甲基-5-异丙基苯酚), 或称香荆芥酚，广泛存在于自然界石香薷、百里香、麝香草、牛至等植物中，具有抗菌、抗炎、抗氧化、增强免疫力和促进生长等多种生物学功效；以香芹酚为主要成分的牛至油（>80%）是重要的食品抗菌防腐保鲜剂，也是我国农业部批准使用的饲料添加剂之一，具有绿色、安全、高效等特点。研究表明香芹酚具有非常强的表面活性和脂溶性，能抑制和清除病原菌生物膜，穿透细胞膜，杀灭病原微生物。斑点叉尾鮰和虹鳟饲料添加香芹酚能促进饵料转化率、抗氧化酶活性和抗病原菌感染能力。有鉴于此，开发香芹酚及其组合物来防治水产动物疾病具有合理性和可行性。目前，尚未见有关香芹酚应用于鱼类粘孢子虫病防治的专利及相关研究。
发明内容
本发明的目的是克服上述不足之处，提供一种用于防治鱼类粘孢子虫病的药物，其在离体条件下能有效杀死粘孢子虫成熟孢子，添加到饲料中投喂能有效预防控制鱼类粘孢子虫病发生。
本发明的技术方案，一种用于防治鱼类粘孢子虫病的药物，包括香芹酚。
作为本发明的一种实施方式，将香芹酚及其药物组合物作为添加剂添加入饲料中。
作为本发明的一种实施方式，所述香芹酚的浓度范围以鱼体重计算，为50-500mg/kg。
作为本发明的一种实施方式，所述药物组合物为具有缓释功能的载体、颗粒、乳化剂或针剂。
作为本发明的一种实施方式，将香芹酚直接制成液体状药物剂型，或以香芹酚为主要成分的牛至油药物。
本发明的另一技术方案是将所述香芹酚及其药物组合物作为主要成分的水产疾病防控药物。
本发明的有益效果：本发明提供的药物在离体条件下能有效杀死粘孢子虫成熟孢子，添加到饲料中投喂能有效预防控制鱼类粘孢子虫病发生，成本低廉，应用效果好。
附图说明
图1 显微镜下洪湖碘泡虫孢子图。
图2 荧光显微镜下洪湖碘泡虫孢子杀灭效果图。
具体实施方式
本发明药物试验部分所用99%纯度的香芹酚购自上海阿拉丁生化科技股份有限公司，洪湖碘泡虫的孢子样品收集自发病异育银鲫的病灶组织。
实施例1
通过percoll密度梯度离心法分离纯化了洪湖碘泡虫成熟孢子，在添加胎牛血清的M199培养基中进行短期培养；香芹酚采用DMSO稀释加入培养基，配置药物浓度0、0.625、0.125、0.500µL/mL各试验组，25℃培养48小时，以SYTOX green nucleic acid stain荧光染料染色在荧光显微镜下计数洪湖碘泡虫孢子存活率。试验结果表明添加药物组杀灭率为0、35%、63%、86%左右。孢子杀灭效果在显微镜下计数见附图1和2；图1为显微镜下洪湖碘泡虫孢子图，图2为荧光显微镜下洪湖碘泡虫孢子杀灭效果图，只有被杀灭的孢子才能呈现荧光。
实施例2
以鲫鱼商品饲料(总脂7%，总蛋白30%)为基础日粮，分别添加0、0.05%、0.1%、0.2%、0.4%、0.8%的香芹酚；健康异育银鲫苗种（30-50g）购自无疫病鲫鱼苗种场，随机设置6组，每组20尾鱼，每天按2.5%投饵量饲喂试验鱼，观察记录鱼死亡96小时，评价药物对异育银鲫安全性。试验结果表明各药物浓度组均无死亡，香芹酚在试验浓度下对鲫鱼具有安全性。
实案例3
在室外2亩养殖池塘中，放入从疫区购买异育银鲫苗种（50-70g）5000尾，随机抽样50尾，解剖取假鳃、卵巢、肾，商品化试剂盒提取组织基因组DNA, 采样洪湖碘泡虫特异PCR检测方法确认放养鱼种携带感染洪湖碘泡虫为40%。在养殖水温25-28℃期间，投喂香芹酚药饵（鲫鱼商品饲料添加0.1%香芹酚）3天，2周后重复投喂药饵1次。在整个养殖周期（8周）内池塘异育银鲫没有发生洪湖碘泡虫病。
Claims (4)
1.香芹酚在制备防治鱼类粘孢子虫病的药物中应用。
2.根据权利要求1所述香芹酚在制备防治鱼类粘孢子虫病的药物中应用，其特征是：将香芹酚及其药物组合物作为添加剂添加入饲料中。
3.根据权利要求1所述香芹酚在制备防治鱼类粘孢子虫病的药物中应用，所述香芹酚的浓度范围以鱼体重计算，为50-500 mg/kg。
4.根据权利要求2所述香芹酚在制备防治鱼类粘孢子虫病的药物中应用，其特征是：所述药物组合物为具有缓释功能的颗粒、乳化剂或针剂。
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